前列腺素F2α和E2对奶牛子宫内膜的修复与保护作用机制研究

Prostaglandins, as a local hormones, which through activating their specific receptors play an important role in female reproductive processes such as ovulation, fertilization and embryo development. But the physiological effect of prostaglandins in endometrum remain undefined. In this research, in order to clarifying the tissue repair and protective effects of prostaglandin F2α and E2 in bovine endometrium, the contents of these experiments are to investigate the effects of prostaglandin F2α and E2 in bovine endometrial epithelial cells expression and secretion of VEGF, CTGF, TGF, IL-8, mucin and β－defensin that are regarded as factors of tissue repair or protection; to invetigate the role of prostaglandin F2α and E2 on endometrial repair and protection; to identify the influence of the LPS-PGE2-EP receptor pathway in prostaglandin mediated endometrial repair and protection; to examine the alteration of VEGF, CTGF, TGF, IL-8, mucin and β－defensin expression in endometrium which isolated from bovine that diagnosed endometritis, by siRNA transfection, Elisa, real time PCR , patch clamp system and immunohitochemical staining. The results from this research will provide theory evidence and experimental evidence for clinical treatment of endometriun-caused infertility by using prostaglandin preparations or non-steroid anti-inflammatory drugs.

前列腺素类化合物作为局部激素通过激活其特定受体在排卵、受精以及胚胎发育等生殖过程中发挥着重要作用，但目前其对子宫内膜的作用机制尚不清楚。本项目为了阐明PGF2α和PGE2对奶牛子宫内膜的修复与保护作用机制，采用siRNA基因沉默技术、膜片钳技术、Elisa法、荧光定量PCR法、免疫组化染色法等，研究PGF2α和PGE2对奶牛子宫内膜上皮细胞合成分泌VEGF、CTGF、TGF、IL-8、β－防御素、黏液素等细胞组织生长或黏膜保护相关因子的作用；研究PGF2α和PGE2对子宫内膜组织修复与保护作用；研究LPS－PGE2－EP受体途径对前列腺素介导的子宫内膜修复与保护的影响；研究患子宫内膜炎病牛的子宫内膜表达COX、VEGF、CTGF、TGF、IL-8、β－防御素等的变化。项目成果可为临床上合理应用前列腺素类药物或非甾体类抗炎药防治奶牛子宫内膜炎性不孕症提供理论依据和实验基础。

前列腺素类化合物作为局部激素通过激活其特定受体在排卵、受精以及受精卵的发育等生殖生理过程中发挥重要作用，但目前其对子宫内膜的作用机制尚不清楚。本研究已经按照预定计划完成了规定的任务，主要在前列腺素E2（prostaglandin E2，PGE2）、PTGER2受体激动剂butaprost和PTGFR受体激动剂fluprostenol对奶牛子宫内膜组织生长因子表达及细胞增殖的影响方面取得了一定的成果。实验结果表明，PGE2、butaprost和fluprostenol能够诱导奶牛子宫内膜组织中结缔组织生长因子（connective tissue growth factor，CTGF）、成纤维细胞生长因子-2（Fibroblast growth factor-2，FGF-2）、白细胞介素-8（interleukin-8, IL-8）、转化生长因子-β（transforming growth factor-β，TGF-β)、基质金属蛋白酶2（matrix metallopeptidase 2，MMP-2）和血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）的表达。此外奶牛子宫内膜组织中增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）、细胞角蛋白18（cytokeratin 18，CK18）、成纤维细胞特异性蛋白-1（fibroblast-specific protein-1，FSP-1）的表达也能够被PGE2、butaprost和fluprostenol所诱导，并且对半胱氨酸天冬氨酸特异性蛋白酶（caspase-3）蛋白的表达具有抑制作用。上述结果说明前列腺素类化合物可能对奶牛子宫内膜组织的修复具有一定促进作用。本课题组的研究成果可为临床上合理应用前列腺素类药物防治奶牛繁殖障碍症并提高奶牛繁殖能力提供理论依据和实验依据。